Leaky gut (decreased intestinal barrier function) decreases milk yield and DMI, and might be related to metabolic disease in dairy cows. Six cannulated Holstein cows (36.3 ± 1 kg/d milk; 212 ± 97 DIM; 691 ± 70 kg BW) were used in a replicated 3 × 3 Latin square design experiment with 21-d periods (d 1–16 wash-out and d 17–21 challenge) to develop a model to induce leaky gut via feed restriction or altered hind gut fermentation. Treatments during the challenge were ad libitum feeding (CTR), 50% feed restriction (FR), and 500 g/d DM resistant starch (RS; abomasally infused once a day before feeding). Blood samples were collected 4 h after feeding on d 13 and 14 to create a baseline for responses measured, and on d 17, 19, and 21 of each period. Fecal samples were collected 4 and 8 h post-feeding on d 14 and 21. Data were analyzed using the GLIMMIX procedure of SAS with fixed effects of treatment, challenge day (repeated measures) and their interaction, and random effects of square, period, and cow. Baseline values were used as covariates. During the challenge period, FR decreased DMI by 48% and ECM yield overtime (25.4 vs. 28.0 kg/d) compared with CTR and RS (both P < 0.05). Overall, FR increased plasma NEFA compared with CTR and RS (160 vs. 82.4 mEq/L; P < 0.05). In addition, FR increased plasma total proteins (7.57 vs. 7.36 g/dL), albumin (3.69 vs. 3.43 g/dL), and γ-glutamyltransferase (37.9 vs. 31.8 U/L) at d 19, and decreased alkaline phosphatase (41.4 vs. 49.1 U/L) at d 21 compared with CTR (all P < 0.05). Responses to the RS treatment were intermediate between CTR and FR for the last parameters mentioned. Treatments did not affect plasma concentrations of lipopolysaccharide binding protein, endotoxin, serum amyloid A, interleukin 1-β, and serum coloration (all P > 0.20). Treatments did not affect fecal pH 4 h post-feeding. However, RS decreased fecal pH 8 h post-feeding compared with CTR and FR (6.21 vs. 6.80; P < 0.05). Lack of responses on inflammatory biomarkers measured in this experiment suggests the models tested did not affect intestinal barrier function, and therefore, a more intense challenge may be needed to induce leaky gut in healthy post-peak cows. Further, evaluating these insults earlier in lactation is of scientific and practical interest.